Original paper Acrodermatitis chronica atrophicans: various faces of the late form of Lyme borreliosis Anna Moniuszko-Malinowska, Piotr Czupryna, Justyna Dunaj, Sławomir Pancewicz, Adam Garkowski, Maciej Kondrusik, Sambor Grygorczuk, Joanna Zajkowska Department of Infectious Diseases and Neuroinfections, Medical University of Białystok, Białystok, Poland Adv Dermatol Allergol 2018; XXXV (5): 490–494 DOI: https://doi.org/10.5114/ada.2018.77240 Abstract Introduction: Acrodermatitis chronica atrophicans (ACA) is probably the most common late and chronic manifesta- tion of the Lyme borreliosis seen in European patients. Aim: To analyze epidemiological data, and to investigate the effects of treatment of patients with ACA. Material and methods: Nine patients were included in the study. All patients had serological examinations (ELISA and Western blot) and histopathological examination of the skin lesions performed. Eight patients had PCR in the skin biopsy performed. Results: The duration of symptoms ranged from 2 months to 2 years. In 7 patients, skin lesions were located on lower limbs, in 2 patients – in a non-typical body area – abdomen. In 1 patient, scleroderma and in 3 patients, diabetes mellitus was diagnosed. Borrelia burgdorferi DNA was detected in 25% of the skin biopsy specimens. IgG anti-B. burgdorferi specific antibodies were present in serum of all patients (confirmed by Western blot). In all cases, the diagnosis was confirmed by histopathological examination. The response to ceftriaxone therapy varied. In 5 cases, the lesions resolved completely, in others they faded. Conclusions: Despite raising awareness of Lyme borreliosis, late forms of the disease such as ACA are still observed. Acrodermatitis chronica atrophicans skin lesions may be located in non-characteristic areas, e.g. abdominal skin. Symptoms are not irritating or painful, therefore patients do not seek medical help. The effect of antibiotic treat- ment varies. Key words: acrodermatitis chronica atrophicans, Lyme borreliosis. Introduction 0.3% of LB cases [5]. In Norway, ACA accounts for 5% of Lyme borreliosis (LB) in Europe is caused by a geno- all clinical cases of LB [6], and in northern Italy – 2.5% [7]. species of the Borrelia burgdorferi sensu lato complex. The diagnosis of ACA is much more difficult than that There are three skin manifestations of LB: erythema mi- of EM or BL as the clinical manifestations of ACA may grans (EM), borrelial lymphocytoma (BL), and acroderma- vary. Acrodermatitis chronica atrophicans is probably the titis chronica atrophicans (ACA) [1]. most common late and chronic manifestation of the LB In Europe, LB with all its dermatologic manifestations seen in European patients. occurs in almost all countries, predominantly in the cen- tral part of the continent. The annual incidence ranges from 9.4 cases per 100,000 population in France to Aim 120 cases per 100,000 population in north-eastern Po- The objectives of the study were to verify human land, 130 cases per 100,000 population in Austria and 155 cases per 100,000 population in Slovenia [2–4]. cases of ACA, to analyze their epidemiological data, and The overall prevalence of ACA in all European pa- to investigate the effects of treatment of patients from tients with LB is about 1–10%, depending on the region. the Podlaskie region of Poland, considered as endemic For example, in Bulgaria, both BL and ACA account for for tick-borne diseases. Address for correspondence: Anna Moniuszko-Malinowska, Department of Infectious Diseases and Neuroinfections, Medical University of Białystok, 14 Żurawia St, 15-540 Białystok, Poland, phone: +48 85 740 95 14, fax: +48 85 740 95 15, e-mail: [email protected] Received: 24.04.2017, accepted: 17.08.2017. 490 Advances in Dermatology and Allergology 5, October / 2018 Acrodermatitis chronica atrophicans: various faces of the late form of Lyme borreliosis Material and methods bromide (5 µg/ml; Syngen, USA) at 80 V for 80 min. The Nine patients (5 women, 4 men; mean age: 58.7 ±14.3 results of the PCR were viewed under UV light (UV to years-old, 4 (44%), inhabitants of cities) with ACA were Gel Logic System 100 (Kodak Imaging System, Inc., USA). included in the study. All patients were tested for anti- Probes with the PCR product of the size of 276 base pairs B. burgdorferi antibodies of the IgM and IgG class by ELI- (bp) were regarded positive. The internal control had SA (Borrelia recombinant IgG and IgM High Sensitivity, a size of 420 bp. For precise detection of B. burgdorferi Biomedica, Austria). Results > 11 BBU/ml in the IgM and s.l. fla, amplicons and internal control molecular weight IgG classes were considered positive. marker (M100-500-Blirt S.A. Poland) was used. The studies have been approved by the ethics com- mittee of the Medical University of Bialystok, Poland. Results In all cases, results were confirmed by Western blot testing (Biomedica, Austria). In all patients, the his- Eight (88%) of our patients remembered a tick bite. topathological examination of skin samples was per- Five (55%) patients had been previously treated with an- formed. tibiotics because of EM. In 5 (56%) cases, this was a job- From 8 patients, skin biopsy samples of 3 mm in di- related disease. The duration of symptoms ranged from ameter were taken and examined for the presence of 2 months to 2 years. In 7 (77%) patients, skin lesions B. burgdorferi s.l. DNA by nPCR (nested PCR). After collec- were localized on the lower limbs. Two patients had co- tion, skin biopsy samples were placed in aseptic round- existing skin lesions in a non-typical body area – abdo- ed-bottom tubes and frozen or prepared fresh. All skin men (Figures 1–6). Six (66%) patients suffered from mus- biopsies were cut into smaller parts with stainless steel cle and 6 (66%) from joint pain. In 1 patient, scleroderma knife-edge and covered with a maximum of 80 µl PBS and in 3 (33%) patients, diabetes mellitus was diagnosed buffer. as concomitant diseases. In all patients, ELISA in the IgG class was positive DNA extraction (mean titer: 90.5 ±9.1 BBU/ml). All results were confirmed DNA from skin biopsies was extracted by using a Qia- by Western blot. Histopathological examination revealed gen DNeasy Blood and Tissue Mini kit according to the features typical of ACA: infiltration of lymphocytes, his- manufacturer’s instructions. Purified DNA (100 µl) was tiocytes and plasma cells, prominent vascular channels then frozen at –20°C before amplification. together with telangiectasia, atrophy of the dermis and subcutis, epidermal atrophy and loss of rete ridges, PCR protocol for Borrelia burgdorferi s.l. which confirmed the diagnosis. Borrelia burgdorferi s.l. DNA was detected in 25% of A fragment of the flagene, the specific DNA sequence the skin biopsy specimens. Detailed characteristics of encoding flagellin was used for B. burgdorferi s.l. detec- tion in “one tube” nested PCR. The Borrelia burgdorferi patients is presented in Table 1. PCR kit (GeneProof, Czech Republic) for in vitro diagnos- Laboratory tests (blood morphology, aminotransfer- tics was used for this purpose, minimizing non-specific ases activity, creatinine concentration, C-reactive protein reactions and maximizing sensitivity because of the “hot (CRP) concentration) revealed no abnormalities. start” technology employed. Possibility of PCR inhibition All patients were treated with third-generation cephalo- is prevented by addition of internal control into the reac- sporin (ceftriaxone 2 g/day) for 28 days (according to Polish tion mix. The risk of contamination is prevented by using guidelines). The outcome of treatment varied. In 5 cases, uracil-DNA-glycosylase (UDG). 4 µl of the template DNA the changes disappeared completely (after 1 to 6 months), isolates was added to 36 µl of the MasterMix for final in others faded, but skin thinning and discoloration of bluish reaction mix volume of 40 µl. The course of the reaction pink remained (Table 1). No side effects of antibiotic therapy in accordance with the manufacturer’s instructions was were observed. performed on the SensoQuest LabCycler (SensoQuest, Germany) with authors’ own modifications. Nested PCR Discussion was performed according to the following amplification program: UDG decontamination, initial denaturation at Acrodermatitis chronica atrophicans is a late and 96°C for 10 min, first amplification for 30 cycles (denatur- long-lasting form of LB, which may be present for many ation at 96°C for 20 s, annealing at 68°C for 20 s, exten- years. It is characterized by red or bluish-red lesions and sion at 72°C for 40 s), second amplification for 45 cycles leads to extensive flaccid atrophy of the skin, which be- (denaturation at 96°C for 20 s, annealing at 54°C for comes more and more prominent. Fibroid nodules may 20 s, extension at 72°C for 30 s) and final extension at develop over bony prominences and sclerodermic le- 72°C for 2 min. The samples were cooled at +4°C. sions may develop in atrophic skin areas [8]. It may limit The PCR products were separated on 2% agarose gel movement of upper and lower limb joints. ACA is most (Sigma-Aldrich, Germany) with the addition of ethidium commonly located on extremities, although it may affect Advances in Dermatology and Allergology 5, October / 2018 491 A. Moniuszko-Malinowska, P. Czupryna, J. Dunaj, S. Pancewicz, A. Garkowski, M. Kondrusik, S. Grygorczuk, J. Zajkowska Figure 1. Patient 2. Lesions typical of ACA on the left thigh Figure 2. Patient 3. Lesions typical of ACA on the right palm Figure 3. Patient 4. Lesions typical of ACA in the left knee Figure 4. Patient 7. Lesions typical of ACA on the left shank area Figure 5. Patient 8. Lesions typical of ACA on the whole right lower limb other skin areas, such as the face [9].